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PubMed1998

Recombinant DNA techniques for bioremediation and environmentally-friendly synthesis

A number of new recombinant DNA techniques have been developed for genetically engineered microorganisms for biodegradation of environmental contaminants or for the synthesis of small molecules. These techniques include new expression vectors to carry the heterologous genes into the host organism, new mechanisms to control gene expression, containment mechanisms to control persistence of genetically-engineered microorganisms, application of site-directed and random mutagenesis to increase the substrate range or activity of biodegradative enzymes, and methods to track genetically-engineered microorganisms.

Journal Article↗
PubMed1978

Microbial catabolism, the carbon cycle and environmental pollution.

The establishment of a carbon cycle was a necessary prerequisite for the evolution of higher forms of life. This could not have been achieved without the direct participation of oxygen gas in certain metabolic reactions. The controlled activation of oxygen is catalyzed by microbial oxygenases; in principle, activated oxygen is hazardous to all living forms but without it, the degradative segment of the carbon cycle could not operate. The degradation of aromatic compounds is not an esoteric activity of a few specialized microorganisms. It occurs continuously, accompanied by fixation and cycling of oxygen on a massive scale; but like other global biochemical processes it tends to be neglected in general biological curricula. However, knowledge of the scope and limitations of microbial catabolic enzymes is central to the development of rational approaches to many of society's environmental concerns.

Acinetobacter↗
PubMed2026

Microbial aerobic degradation of 4-isopropylnitrobenzene by Sphingobium yanoikuyae strain SG1.

4-Isopropylnitrobenzene (4-IPNB) is a nitroaromatic compound commonly employed as an intermediate in pesticide synthesis and chemical manufacturing. Despite its potential environmental persistence and ecological risks, the microbial degradation pathway of 4-IPNB remains largely unknown. In this study, a Gram-negative bacterium, designated Sphingobium yanoikuyae strain SG1, was isolated from a pesticide manufacturing site in Brazil for its ability to utilize 4-IPNB as the sole added source of carbon, nitrogen, and energy. Aerobic degradation of 4-IPNB by strain SG1 was accompanied by nitrite release, and intermediate-trapping experiments revealed the transient accumulation of 4-isopropylcatechol (4-IPC). Together, these findings support the initial conversion of 4-IPNB to 4-IPC through oxidative denitration and dihydroxylation. Genomic and transcriptomic analyses further inferred several candidate nitroarene dioxygenases that may catalyze this initial reaction. Furthermore, the downstream metabolism of 4-IPC proceeded via both meta- and ortho-cleavage pathways, with cell-extract enzyme assays demonstrating predominant meta-cleavage activity under the tested conditions. Strain SG1 also degraded 4-IPNB in nonsterile soil slurry microcosms, extending its degradation capability beyond defined liquid culture. This study provides the first comprehensive insight into the microbial aerobic degradation of 4-IPNB, advances our understanding of the environmental fate of emerging nitroaromatic contaminants, and supports the potential of strain SG1 in 4-IPNB biodegradation and removal.

Sphingomonadaceae↗
PubMed2005

Toxicology and fate of Pestanal and commercial propetamphos formulations in river and estuarine sediment.

To quantify the impact of organophosphate pesticides on aquatic ecosystems requires a mechanistic understanding of their behaviour in a range of environmental matrices. The objective of this study was to compare the sorption/desorption, biodegradation and toxic effects of the Pestanal grade and commercial formulation (Ectomort Centenary) of the organophosphate insecticide propetamphos in river and estuarine sediments. For both formulations, the sorption of propetamphos onto sediment was initially very rapid followed by a slower sorption phase. Similarly, the initial rate of desorption was rapid, followed by a much slower rate. In both sorption and desorption experiments, the level of sorbed propetamphos was considerably higher for the commercial formulation of propetamphos (Kd=7-11) than for the Pestanal grade (Kd=4-10). The rate of propetamphos biodegradation was sediment dependent but was most rapid where microbial activity and nutrients were the highest and sorption was the lowest. Propetamphos was more rapidly degraded in sediments under aerobic (t(1/2)=15 d) compared to anaerobic conditions (t(1/2)=19 d). However, no significant difference in the biodegradation rates of the Pestanal grade and commercial formulations of propetamphos were observed. The toxic effect of propetamphos on sediment microbial communities was significantly greater for the commercial formulation than for the Pestanal grade of propetamphos based on EC50 (21 versus 236 microg g(-1)) and EC10 values (0.3 versus 54 microg g(-1)). In conclusion, our results highlight the importance of using commercial pesticide formulations when carrying out ecotoxicological testing.

Adsorption↗
PubMed2006

Towards commercial production of microbial surfactants.

Biosurfactants or microbial surfactants are surface-active biomolecules that are produced by a variety of microorganisms. Biosurfactants have gained importance in the fields of enhanced oil recovery, environmental bioremediation, food processing and pharmaceuticals owing to their unique properties--higher biodegradability, lower toxicity, and effectiveness at extremes of temperature, pH and salinity. However, large-scale production of these molecules has not been realized because of low yields in production processes and high recovery and purification costs. This article describes some practical approaches that have been adopted to make the biosurfactant production process economically attractive: these include the use of cheaper raw materials, optimized and efficient bioprocesses and overproducing mutant and recombinant strains for obtaining maximum productivity. The application of these strategies in biosurfactant production processes, particularly those using hyper-producing recombinant strains in the optimally controlled environment of a bioreactor, might lead towards the successful commercial production of these valuable and versatile biomolecules in near future.

Animals↗
PubMed2006

Confronting the realities of wastewater aquaculture in peri-urban Kolkata with bioeconomic modelling.

Wastewater reuse for aquaculture is a reality in several Asian countries, however, traditional practices face constraints including inadequate or contaminated wastewater inputs and growing concern over health risks. Based on Kolkata and a wastewater flow of 550,000m(3)d(-1), rational and conventional designs for lagoon-based wastewater treatment and reuse through aquaculture were compared using bioeconomic modelling. Outcomes showed the rational design required a larger area than the conventional or traditional systems, but that financial returns, nutrient retention and fish production were higher; gross fish production employing rational and conventional designs was 45,500 and 11,560t, respectively. However, limited land availability and constraints to reconfiguring the existing system make implementation of the rational design unlikely. Findings suggest traditional practices could be enhanced by adopting wastewater treatment prior to reuse, modifying fish production strategies, and monitoring to safeguard health. Bioeconomic modelling constitutes a useful tool in comparing treatment and reuse options, permitting the sensitivity of financial returns to changing costs and recent revisions to WHO guidelines for safe wastewater reuse to be assessed, however, social and environmental consequences demand consideration.

Aquaculture↗
PubMed2001

Distribution and diagenesis of organic and inorganic phosphorus in sediments of the Baltic proper.

Measurements of the distribution of organic and inorganic phosphorus in the sediment have been performed at 10 sites, in the central-northern Baltic proper. Variations in deposition environment and environmental properties, such as redox chemistry and bottom water dynamics, apparently affect the distribution by altering the supply and diagenesis of organic and inorganic phosphorus constituents. The C/P and the N/P ratios of the sediment are highly divergent (higher) from the average Redfield ratio for marine living organisms at all examined sites. This indicates preferential autolytic organic phosphorus degradation and/or (deeper down in the sediment) altered input of terrestrial organic matter. The redox condition seems to affect the degradation efficiency of the organic matter since higher concentrations of C(org) occur at anoxic conditions than at oxic. At two sites significant amounts of C(inorg) have been detected indicating authigenic precipitation of carbonates. Further, authigenic precipitation of phosphate minerals also seems to occur at certain environmental conditions.

Baltic States↗
PubMed2003

Potential availability of heavy metals to phytoextraction from contaminated soils induced by exogenous humic substances.

Effective phytoremediation of soils contaminated by heavy metals depends on their availability to plant uptake that, in turn, may be influenced by either the existing soil humus or an exogenous humic matter. We amended an organic and a mineral soil with an exogenous humic acid (HA) in order to enhance the soil organic carbon (SOC) content by 1% and 2%. The treated soils were further enriched with heavy metals (Cu, Pb, Cd, Zn, Ni) to a concentration of 0, 10, 20, and 40 microg/g for each metal and allowed to age at room temperature for 1 and 2 months. After each period, they were extracted for readily soluble and exchangeable (2.5% acetic acid), plant-available (DTPA, Diethylentriaminepentaacetic acid), and occluded (1 N HNO(3)) metal species. Addition of HA generally reduced the extractability of the soluble and exchangeable forms of metals. This effect was directly related to the amount of added HA and increased with ageing time. Conversely, the potentially plant-available metals extracted with DTPA were generally larger with increasing additions of exogenous HA solutions. This was attributed to the formation of metal-humic complexes, which ensured a temporary bioavailability of metals and prevented their rapid transformation into insoluble species. Extractions with 1 N HNO(3) further indicated that the added metals were present in complexes with HA. The observed effects appeared to also depend on the amount of native SOC and its structural changes with ageing. The results suggest that soil amendments with exogenous humic matter may accelerate the phytoremediation of heavy metals from contaminated soil, while concomitantly prevent their environmental mobility.

Biodegradation, Environmental↗
PubMed1998

Bio-dissolution of spent nickel-cadmium batteries using Thiobacillus ferrooxidans.

In this study, the production of sulphuric acid in bioreactors with Thiobacillus ferrooxidans attached on elemental sulphur was investigated. These bioreactors reached a maximum H+ productivity of 80 mmol kg-1 d-1 of support. This medium was used for the indirect dissolution of spent nickel-cadmium batteries recovering after 93 days 100% of cadmium, 96.5% of nickel and 95.0% of iron. Moreover, recoveries higher than 90.0% were reached when anodic and cathodic materials were directly added to Thiobacillus ferrooxidans cultures with sulphur as the sole energy source. The results presented show an economic and effective method which could be considered the first step to recycle spent and and discarded batteries preventing one of the many problems of environmental pollution.

Biodegradation, Environmental↗
PubMed2002

Development and characterization of a green fluorescent protein-based bacterial biosensor for bioavailable toluene and related compounds.

A green fluorescent protein-based Pseudomonas fluorescens strain A506 biosensor was constructed and characterized for its potential to measure benzene, toluene, ethylbenzene, and related compounds in aqueous solutions. The biosensor is based on a plasmid carrying the toluene-benzene utilization (tbu) pathway transcriptional activator TbuT from Ralstonia pickettii PKO1 and a transcriptional fusion of its promoter PtbuA1 with a promoterless gfp gene on a broad-host-range promoter probe vector. TbuT was not limiting, since it was constitutively expressed by being fused to the neomycin phosphotransferase (nptII) promoter. The biosensor cells were readily induced, and fluorescence emission after induction periods of 3 h correlated well with toluene, benzene, ethylbenzene, and trichloroethylene concentrations. Our experiments using flow cytometry show that intermediate levels of gfp expression in response to toluene reflect uniform induction of cells. As the toluene concentration increases, the level of gfp expression per cell increases until saturation kinetics of the TbuT-PtbuA1 system are observed. Each inducer had a unique minimum concentration that was necessary for induction, with K(app) values that ranged from 3.3 +/- 1.8 microM for toluene to 35.6 +/- 16.6 microM for trichloroethylene (means +/- standard errors of the means), and maximal fluorescence response. The fluorescence response was specific for alkyl-substituted benzene derivatives and branched alkenes (di- and trichloroethylene, 2-methyl-2-butene). The biosensor responded in an additive fashion to the presence of multiple inducers and was unaffected by the presence of compounds that were not inducers, such as those present in gasoline. Flow cytometry revealed that, in response to toxic concentrations of gasoline, there was a small uninduced population and another larger fully induced population whose levels of fluorescence corresponded to the amount of effectors present in the sample. These results demonstrate the potential for green fluorescent protein-based bacterial biosensors to measure environmental contaminants.

Benzene↗
PubMed2005

Genetically modified organisms for the environment: stories of success and failure and what we have learned from them.

The expectations raised in the mid-1980s on the potential of genetic engineering for in situ remediation of environmental pollution have not been entirely fulfilled. Yet, we have learned a good deal about the expression of catabolic pathways by bacteria in their natural habitats, and how environmental conditions dictate the expression of desired catalytic activities. The many different choices between nutrients and responses to stresses form a network of transcriptional switches which, given the redundance and robustness of the regulatory circuits involved, can be neither unraveled through standard genetic analysis nor artificially programmed in a simple manner. Available data suggest that population dynamics and physiological control of catabolic gene expression prevail over any artificial attempt to engineer an optimal performance of the wanted catalytic activities. In this review, several valuable spin-offs of past research into genetically modified organisms with environmental applications are discussed, along with the impact of Systems Biology and Synthetic Biology in the future of environmental biotechnology.

Animals↗